Russell L. Miller

Evidence that N-acetylation regulates the behavioral activity of α-MSH in the rat and human central nervous system

alpha-MSH immunoreactive peptides were fractionated and characterized in rat and human brain and rat pituitary by reversed phase high pressure liquid chromatographic techniques. alpha-MSH and deacetylated alpha-MSH were two major naturally existing peptides in both brain and pituitary gland. Subsequent experiments examined the roles of these two peptides in neuronal function. The alpha-MSH was clearly more effective than deacetylated alpha-MSH in improving performance on a visual discrimination task after intraperitoneal administration and in inducing excessive grooming after intraventricular administration. The difference in behavioral potency may be explained by the fact that alpha-MSH was much more resistant to peptidase degradation than was deacetylated alpha-MSH. N-acetylation of alpha-MSH may be an effective regulatory process for modulating the behavioral potency of the secretory product of alpha-MSH-containing pituitary cells and neurons.

Secretin immunoreactivity in rat and pig brain

Secretin immunoreactivity in rat and pig brain has been identified and characterized utilizing a highly specific radioimmunoassay and fractionation on a high pressure liquid chromatographic system reverse phase column. One immunoreactive peak from each brain extract was observed. Secretin immunoreactivity from rat brain and duodenum coelute, but eluted slightly ahead of the immunoreactivity from pig brain and duodenum and from synthetic porcine secretin. Immunoreactive secretin is widely distributed in the thalamus, hypothalamus and olfactory bulb, cerebral cortex, midbrain, septum, striatum, hippocampus, medulla and pons. The highest concentrations occur in the pineal and the pituitary gland.

Release of alpha-melanocyte stimulating hormone into rat and human cerebrospinal fluid in vivo and from rat hypothalamus slices in vitro

The release of alpha-melanocyte stimulating hormone (alpha-MSH) from central nervous system neurons was investigated and demonstrated in vivo and in vitro. alpha-MSH immunoreactivity in rat and human cerebrospinal fluid (CSF) is comprised of deacetylated alpha-MSH, alpha-MSH and the methionine sulfoxide forms of these peptides. The sulfoxides are formed artifactually upon extraction. alpha-MSH in rat CSF is unaffected by hypophysectomy but is markedly increased by electrical stimulation of the mesencephalic central gray. These data indicate that CSF alpha-MSH is primarily of neuronal origin, alpha-MSH is also released in a calcium dependent manner from hypothalamic slices in vitro. The fact that the release of alpha-MSH is stimulated by veratridine and inhibited by tetrodotoxin demonstrates the necessity for neuronal sodium influx for alpha-MSH release. The presence of an alpha-MSH neurosecretory process supports a neurotropic role for this peptide in the central nervous system.

Determination of rat pineal gland melatonin content by a radioimmunoassay.

Abstract Melatonin content in individual rat pineal glands was measured by radioimmunoassay (RIA). The RIA used can very reliably detect as little as 50 pg of melatonin. The various precursors, analogues, and the metabolite of melatonin (6-hydroxymelatonin) which were tested for cross-reactivity were not recognized by the antibody. The effects on melatonin levels in rat pineal glands following the administration of L-tryptophan, 5-hydroxy-L-tryptophan, serotonin, N-acetylserotonin, melatonin and pargyline are also presented.

Secretin receptors in the rat kidney: Adenylate cyclase activation and renal effects

In previous studies it has been demonstrated that pharmacological administration of secretin can alter urine output. Whether the effect is due to a direct action on kidney was investigated by examining the effect of secretin on renal output, and determining whether there were secretin receptors and a secretin sensitive adenylate cyclase in the kidney. Secretin had an antidiuretic action on kidney when administered intravenously to anesthetized hydrated rats. In addition, binding sites for (125I)-secretin, and a secretin sensitive adenylate cyclase were identified in rat kidney. Binding was saturable and reversable and was half maximally inhibited by 1 X 10(-7) M synthetic porcine secretin. Autoradiographic studies revealed a high density of secretin binding sites in the outer medulla of the kidney, a region that is composed mainly of the thick ascending limb of the loop of Henle, and is also the major site of action for the antidiuretic hormone, vasopressin. The data indicate that a functional secretin receptor system exists in kidney which may have a physiological role in regulating urine output.

Radioimmunoassay of pyridostigmine in plasma and tissues

A sensitive and specific radioimmunoassay is reported for the cholinergic agent, pyridostigmine. Antibodies were raised in rabbits against a conjugate of pyridostigmine and bovine serum albumin. The assay can detect as little as 2.5 ng/ml of drug directly in plasma and tissue homogenates. Structurally similar compounds and major metabolites are not recognized by the antibody. The specificity of the antibody has been confirmed by utilizing high pressure liquid chromatography. Plasma concentration-time profiles and tissue distribution of the drug were determined by this method in rat after intramuscular administration of pyridostigmine.